Retroactivity, modularity, and insulation in synthetic biology circuits

Abstract

A central concept in synthetic biology is the reuse of well-characterized modules. Modularity simplifies circuit design by allowing for the decomposition of systems into separate modules for individual construction. Complex regulatory networks can be assembled from a library of devices. However, current devices in synthetic biology may not actually be modular and may instead change behavior upon interconnections, a phenomenon called retroactivity. Addition of a new component to a system can change individual device dynamics within the system, potentially making timeconsuming iterative redesign necessary. Another need for systems construction is the ability to rapidly assemble constructs from part libraries in a combinatorial, highthroughput fashion. In this thesis, a multi-site assembly method that permits the rapid reshuffling of promoters and genes for yeast expression is established. Synthetic circuits in yeast to measure retroactivity and to act as an insulator that attenuates such effect are designed and modeled.